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Description
Human trypsin ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: 1. Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 3. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with trypsin capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of trypsin in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Trypsin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Trypsin is a serine protease from the PA superfamily found in the digestive systems of many vertebrates, where it hydrolyzes proteins. Trypsin is formed in the small intestine and becomes trypsin when its proenzyme form, trypsinogen produced by the pancreas, is activated. It primarily cleaves peptide chains at the carboxyl side of the amino acids lysine or arginine. Trypsin is used in many biotechnological processes. This process is often referred to as trypsinization, and proteins digested/treated with trypsin are said to be trypsinized. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
Based on 1749 reviews
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Product Reviews
★★★★★ 5
Hallelujah… finally, I found the right pillow!!!
Size: Queen (Pack of 1), Color: White
I have spent a lot of money searching for the right pillow. I recently tried to Tempur-pedic that cost twice with this pillow cost. Tempur-pedic flattened out. I’ve had this pill for a few weeks now and it works wonders. I am a paraplegic. You can have a lot of neck problems. I love the fact that this pillow is adjustable. I immediately saw it was too stuffy and took a bunch out. It works perfect so glad I didn’t spend money on a $200 purple pillow.
I read a lot of reviews and there were a lot of complaints about the odor. Mine did smell funny when it came out of the box. I immediately threw it in the dryer with a few bounce fabric sheets came out smelling fine.
Pillows are a very individual thing, but I think for 40 bucks this pillow is worth a try. I’m really enjoying mine about one of those silk pillowcases to go over it and it sleeps cool and comfortable. Best wishes in your search for the right pillow.
Update: I’ve been using this pillow several months and it continues. Give me a good night rest pain-free.. well worth the money.
UPDATE: I have been using this pillow for three months and it still works great. It does not flat out or get hot..
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 29, 2026
★★★★★ 5
Soft comfortable very good pillow
Size: King (Pack of 1), Color: White
I really love my new pillow it's great and comfortable. Yes, it has a little smell, but if you wash the pillowcase and put the pillow on the window for a day you won't notice anything anymore.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 18, 2026
★★★★★ 5
Best place for head
Size: Queen (Pack of 1), Color: White
Do you love not getting sleep and sweating all over your pillow at night when you are asleep? If so then don't buy this pillow! If you want a good nights sleep and cool pillow like me this is the one. Ive always been ehh towards spending so much on a pillow but its 100% worth it, I sleep so much better and it forms to my head and keeps me cool at night. Great support even for my big head and comes with a bag of extra stuffing if you want to have a little more firmer pillow. Highly recommend
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Reviewed in the United States on April 4, 2026
★★★★★ 3
Oversized
Size: Queen (Pack of 2), Color: White
I have chronic neck pain. I am forever buying new pillows. Sometimes I think I found the perfect pillow but then three months later they’re losing their loft. I thought I would give these pillows to try since the stuffing is removable. Now these pillows are really oversized. To the point where I thought that they had sent me king-size pillows but I double checked the packaging and they were Queen. I ended up having to buy king-size pillowcases for them because they just barely fit in the queen size and the pillowcases would always end up pulling back from the edge. So that’s a negative one star there. Now I’ve been sleeping on these pillows for a while and they have held up well. I did end up removing some of the stuffing because it was just too much. I’m a side sleeper and when I use this pillow, it does a good job of keeping my neck in alignment. However, the pillow tends to migrate up as I sleep. Or I tend to slip down off the pillow, either or. So that’s why I’ve removed a second star. Maybe if I removed more stuffing that wouldn’t happen, but then I feel like the loft would be too little. I will say that these provide great support when sitting up in bed reading or watching TV. So would I buy them again? Probably not. Am I going to keep using them? Yes
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Reviewed in the United States on December 23, 2025
★★★★★ 5
Amazing - Stop Scrolling and Buy This Pillow
Size: Queen (Pack of 1), Color: White
I have been an Amazon customer ever since the very beginning, and this is my first review. Not because I haven’t purchased tons of amazing things (and a few bad ones), just never felt the need.
However, I researched pillows for longer than I care to admit, and decided on this one.
I’ve had it for about six months and absolutely love it. No lie, I genuinely look forward lying my big head on it after a long day.
I am a side sleeper, but flip flop all around through the night and it’s great in every position.
Pros:
Stays nice and cool, which is soothing regardless of the time of year.
Has a sort of quilted textured pattern on the zip up cover, gives it a luxury feel, that breaths nicely even when a pillow case is added.
The fill is fully adjustable to your preference.
I knew it came with extra filling but was worried it would not be enough. Wrong. It was very full and I ended up taking some out until it was perfect.
It comes vacuum sealed and suggests it may take some time to fully take shape. However, I found it to be about 90% almost instantly and was sleeping on it a couple hours later.
There was no unpleasant scent sometimes associated with memory foam items.
The foam itself is very nice, a mix of what appears to be shredded and cotton candy cloud like. Very brilliant idea, as it is somehow supportive, soft and firm all at once.
The affordability of this pillow compared to some of the others in this category can’t be overstated.
Oh, and the packaging was very thoughtfully and securely done.
If you are like me, an over thinker, I would suggest just taking this for a spin. It’s amazing.
I’m not a bot, or working for the company. I’m just truly satisfied, love this pillow, and have been sleeping great.
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Reviewed in the United States on April 6, 2026
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